Mass Production of Beneficial Organisms: Invertebrates and Entomopathogens

Front Cover
Mass Production of Beneficial Organisms
Copyright Page
Dedication
Contents
List of contributors
Preface
Reference
Section I
1 Introduction
1.1 Challenges of mass-producing beneficial organisms
1.2 Challenges of arthropod mass production for biological control
1.3 Challenges of mass-producing pathogens for biological control
1.4 Challenges of mass-producing invertebrates for their products and ecological services
References
Further reading
2 Production of coleopteran predators
2.1 Introduction
2.1.1 Aims of this chapter
2.1.2 Predatory beetles in culture
2.1.3 Overview of the content
2.2 Foods and production of predators
2.2.1 Feeding preferences and natural prey
2.2.2 Feeding on factitious foods and plant products
2.2.3 Feeding on artificial diets
2.3 Rearing density and production
2.3.1 Crowding
2.3.2 Cannibalism
2.3.3 Design of oviposition substrates and rearing enclosures
2.3.4 Rearing scale
2.4 Temperature and production
2.4.1 Optimizing temperature for rearing
2.4.2 Reducing temperature for cold storage
2.5 Quality control and production
2.5.1 Safeguards against unwanted pathogens and parasites
2.5.2 Preventing colony deterioration
2.5.3 In-shipment, postshipment and prerelease assessments
2.6 Conclusions and recommendations
2.6.1 Synthesis
2.6.2 Future research
Acknowledgments
References
3 Production of heteropteran predators
3.1 Introduction
3.2 Foods
3.2.1 Natural prey
3.2.2 Factitious prey
3.2.3 Artificial diets
3.2.3.1 Definitions
3.2.3.2 Effect on development and reproduction
3.2.3.3 Effect on predation potential
3.2.3.4 Challenges for the practical use of artificial diets
3.3 Plant materials and alternatives
3.3.1 Plant substrates
3.3.2 Artificial substrates
3.4 Abiotic conditions
3.4.1 Optimal climatic conditions for rearing
3.4.2 Cold storage
3.5 Crowding and cannibalism
3.6 Microorganisms
3.7 Breeding and colony maintenance
3.8 Mass-rearing systems
3.9 Conclusion
Acknowledgments
References
4 Production of dipteran parasitoids
4.1 Introduction
4.2 Dipteran parasitoids as biocontrol agents
4.2.1 Tachinidae
4.2.2 Other dipteran parasitoids
4.2.3 Side effects
4.3 Aspects of dipteran parasitoid biology of special interest for production
4.3.1 Host range
4.3.2 Oviposition strategies
4.3.3 Host–parasitoid interactions
4.4 Production techniques
4.4.1 In vivo production
4.4.1.1 Larval food: natural hosts versus alternative hosts
4.4.1.2 Infestation mode
4.4.1.3 Influence of host age at parasitization
4.4.1.4 Abiotic conditions for preimaginal development
4.4.2 In vitro production
4.4.2.1 Nutritional requirements and other needs of dipteran parasitoids
4.4.2.1.1 Nitrogen sources
4.4.2.1.2 Lipids
4.4.2.1.3 Carbohydrates
4.4.2.1.4 Miscellaneous
4.4.2.1.5 Other needs
4.4.2.2 In vitro rearing
4.4.2.3 Continuous in vitro culture
4.4.3 Adult maintenance
4.4.3.1 Food and water supply
4.4.3.2 Space availability and adult management
4.4.3.3 Abiotic conditions
4.4.4 Sterilization and antimicrobial agents
4.4.5 Quality control
4.4.6 Storage and shipment procedures
4.5 Perspectives and concluding remarks
References
5 Production of hymenopteran parasitoids
5.1 Introduction
5.2 Mass rearing of aphelinid parasitoids of the silverleaf whitefly
5.3 Laboratory culture
5.3.1 Plant culture
5.3.2 Whitefly oviposition
5.3.3 Parasitoid culture
5.4 Outdoor field cage production
5.4.1 Plant culture
5.4.2 Whitefly oviposition
5.4.3 Parasitoid culture
5.5 Large-scale greenhouse-based system
5.5.1 Plant production
5.5.2 Insect and disease control
5.5.3 Whitefly colony
5.5.4 Parasitoid production
5.5.5 Parasitoid processing
5.5.6 Storage of parasitoid pupae
5.6 Final remarks
5.7 Production of Tamarixia radiata Watson parasitoid of Diaphorina citri Kuwayama
5.8 Diaphorina citri
5.8.1 Taxonomy
5.8.2 Origin and distribution
5.8.3 Ecology and habits
5.9 Tamarixia radiata
5.9.1 Taxonomy
5.9.2 Origin and distribution
5.9.3 Ecology and habits
5.10 Mass production
5.10.1 Infrastructure, equipment, and materials
5.11 Host plant production
5.11.1 Characteristics, advantages, and disadvantages of using Murraya paniculata
5.11.2 Seed collection
5.11.3 Pulping and transport of fruit
5.11.4 Seed drying and storage
5.12 Production of Murraya paniculata
5.12.1 Substrate preparation
5.13 Sowing
5.13.1 Transplanting and watering
5.13.2 Fertilization
5.13.3 Pruning
5.13.4 Uses and reuse of plants
5.14 Host insect production
5.14.1 Environmental conditions for rearing
5.14.2 Selection of adults for reproduction
5.15 Parasitoid production
5.15.1 Obtaining broodstock
5.15.2 Environmental conditions for breeding
5.15.3 Parasitization
5.15.4 Collection of adults
5.15.5 Handling and packaging prior to release
5.16 Breeds of Tamarixia radiata established in other countries
5.17 Production of parasitoids of muscoid flies
5.18 Host production
5.19 Parasitoid rearing and housing
5.19.1 Host:parasitoid ratios
5.19.2 Use of killed host pupae for parasitoid production
5.19.3 Disease concerns
5.20 Production of Catolaccus grandis (Burks) parasitoid of the boll weevil
5.20.1 In vivo production
5.20.2 Factitious hosts
5.20.3 In vitro production
5.21 Final remarks and future perspective
USDA disclaimer
References
Further reading
6 Mass-production of arthropods for biological control of weeds: a global perspective
6.1 Introduction
6.1.1 Theory and rationale for biological control of weeds
6.1.2 Scope of chapter
6.2 Scope of mass-rearing of biological control agents of weeds
6.2.1 Definition of mass-rearing of weed biological control agents
6.2.2 Summary of the extent of use of mass-rearing in weed biological control
6.2.3 Benefits of mass-rearing in biological weed control
6.3 Critical factors in the design and use of mass-rearing protocols in biological weed control
6.3.1 Decision-making regarding the need for mass-rearing
6.3.2 Decision-making regarding the feasibility of mass-rearing
6.3.3 Critical factors in the design of mass-rearing protocols
6.3.3.1 Production of host substrates
6.3.3.2 Knowledge of agent population biology and plant-insect ecology
6.3.4 Implementation of mass-rearing
6.3.4.1 Monitoring of output level and efficiency
6.3.4.2 Evaluation of success of mass-rearing
6.4 Case studies on mass-rearing in biological weed control
6.4.1 United States
6.4.1.1 History of use
6.4.1.2 Case study: a shoot tip-galling wasp on arundo
6.4.1.2.1 Key challenges in designing a mass-rearing program
6.4.1.2.2 Summary of the mass-rearing protocol
6.4.1.2.3 Summary of output of mass-rearing and cost
6.4.1.2.4 Impact of mass-rearing on agent establishment and efficacy
6.4.2 Canada
6.4.2.1 History of use
6.4.2.2 Case study: a root weevil on houndstongue
6.4.2.2.1 Key challenges in designing the mass-rearing program
6.4.2.2.2 Summary of the mass-rearing protocol
6.4.2.2.3 Output of mass-rearing and estimated cost
6.4.2.2.4 Impact of mass-rearing on establishment and efficacy
6.4.3 South Africa
6.4.3.1 History of use
6.4.3.1.1 History of mass-rearing in biological control
6.4.3.1.2 Mass-rearing as employment opportunities
6.4.3.1.3 Mass-rearing programs at schools
6.4.3.2 Case study: a leaf-feeding planthopper on waterhyacinth
6.4.3.2.1 Key challenges in designing mass-rearing program
6.4.3.2.2 Output of mass-rearing and estimated cost
6.4.3.2.3 Impact of mass-rearing on establishment and efficacy
6.4.4 Australia
6.4.4.1 History of use
6.4.4.2 Case study: two leaf-feeding moths on Parkinsonia
6.4.4.2.1 Key challenges in designing mass-rearing program
6.4.4.2.2 Summary of the mass-rearing protocol
6.4.4.2.3 Output of mass-rearing and estimated cost
6.4.4.2.4 Impact of mass-rearing on establishment and efficacy
6.4.5 New Zealand
6.4.5.1 History of use
6.4.5.2 Case study: a leaf-feeding butterfly on Japanese honeysuckle
6.4.5.2.1 Key challenges in designing mass-rearing program
6.4.5.2.2 Brief summary of mass-rearing protocol
6.4.5.2.3 Output of mass-rearing, estimated cost, and impact
6.4.5.3 Case study: a leaf-feeding beetle on Tradescentia
6.4.5.3.1 Key challenges in designing mass-rearing program
6.4.5.3.2 Brief summary of mass-rearing protocol
6.4.5.3.3 Output of mass-rearing and estimated cost
6.4.5.3.4 Impact of mass-rearing on establishment and efficacy
6.5 Summary and conclusions
6.5.1 Conclusions from case studies
6.5.1.1 Factors that prompted mass-rearing
6.5.1.2 Factors conducive to successful mass-rearing
6.5.1.3 Costs of mass-rearing
6.6 Recommendations
6.6.1 Measuring and communicating benefits of mass-rearing
6.6.2 Keeping mass-rearing at the forefront of implementation
6.6.3 Frontiers in mass-rearing of weed biological control agents
Acknowledgments
References
7 Mass production of predatory mites: state of the art and future challenges
7.1 Introduction
7.1.1 Mites and their importance as biocontrol agents
7.1.2 Brief historical overview
7.2 Phytoseiidae
7.2.1 Lifestyles of phytoseiid predatory mites
7.2.2 Mass-rearing systems for phytoseiid predatory mites
7.3 System 1: both tetranychid prey mites and predatory mites are produced on plants in greenhouses
7.4 System 2: tetranychid prey mites are reared on plants in greenhouses. The predator is reared in climate rooms on detach...
7.5 System 3: tetranychid prey mites are reared on plants in greenhouses. The predator is reared in a climate room on pure ...
7.6 System 4: predatory mites are grown on factitious food sources
7.6.1 Factitious prey mites
7.6.2 Other factitious food
7.7 System 5: predatory mites grown on plants or parts thereof using pollen
7.8 System 6: predatory mites are grown on artificial diet
7.8.1 From laboratory colony to mass production scale: a huge step
7.9 Prey mite
7.9.1 Suitable species
7.9.2 Suitable life stages
7.9.3 Suitable diet/rearing substrate
7.9.4 Predator:prey ratio
7.10 Climate management
7.10.1 Carbon dioxide concentration
7.10.2 Temperature and metabolic heat
7.10.3 Relative humidity and substrate moisture content
7.11 Intraspecific competition
7.12 Contamination management
7.13 Nonphytoseiid predatory mites
7.13.1 Soil predatory mites
7.13.2 Poultry mite predators
7.13.3 Prostigmatid predators
7.14 Diseases
7.14.1 Spider mites—the case of Neozygites
7.14.2 Astigmatid prey mites
7.14.3 Predatory mites
7.15 Challenges and future prospects
7.15.1 Off-plant rearing systems for types I and IV predatory mites
7.15.2 Artificial diets
7.15.3 Role of endosymbionts
7.15.4 Automation
7.15.5 Strain selection
7.15.6 Genetic variation
References
8 Artificial diet development for entomophagous arthropods
8.1 Introduction
8.1.1 Levels of development
8.1.2 Degrees of difficulty
8.2 Arthropod nutrition
8.2.1 Carbohydrate
8.2.2 Lipid
8.2.3 Protein
8.2.4 Vitamins
8.2.5 Minerals
8.3 Determining the basic diet formulation
8.3.1 Chemical analysis
8.3.2 Water content
8.3.3 Macronutrient ratios
8.4 Presentation
8.4.1 Feeding adaptations
8.4.2 Encapsulation of liquid diets
8.4.3 Gels and carriers for solid formulations
8.4.4 Feeding stimulants
8.5 Diet refining
8.5.1 Improving diet quality
8.5.2 From chemically defined to economically sound
8.5.3 Industrialized insect components
8.5.4 Dietary self-selection
8.5.5 Diet preservation
8.6 Future perspectives
8.6.1 Multiple diet component testing
8.6.2 Multiomic assessment of diet quality
8.6.3 Endosymbionts
8.7 Concluding remarks
References
9 Concepts and methods of quality assurance for mass-reared parasitoids and predators
9.1 Introduction
9.2 Quality assurance in the marketplace
9.3 Customer involvement in quality assurance
9.4 Building a complete quality assurance system
9.4.1 Management
9.4.2 Methods development
9.4.3 Material
9.4.4 Production
9.4.5 Research
9.4.6 Utilization
9.4.7 Personnel
9.4.8 Quality control
9.5 Quality assessments of mass-reared natural enemies
9.6 Quality assurance and control data acquisition and analysis
9.7 Quality assurance system review
9.7.1 Approach
9.7.2 Review of functions (successes and failures)
9.7.3 Conclusions
9.7.4 Recommendations (based on evidence and insights)
9.7.4.1 Attachments to report
9.8 Research on quality assessment for mass-reared parasitoids and predators
9.9 Conclusion
Acknowledgements
References
Section II
10 Production of entomopathogenic nematodes
10.1 Introduction
10.2 In vivo production
10.2.1 Basic method
10.2.2 Factors affecting efficiency
10.2.3 Recent advances and future directions
10.3 In vitro production—solid culture
10.3.1 Basic method
10.3.2 Factors affecting efficiency
10.3.3 Recent advances and future directions
10.4 In vitro production–liquid culture
10.4.1 Basic method
10.4.2 Factors affecting efficiency
10.4.3 Recent advances and future directions
10.5 Analysis and conclusion
10.5.1 Comparison of production methods
10.5.2 Strain selection, improvement and stability
10.6 Conclusion
References
11 Mass production of entomopathogenic fungi—state of the art
11.1 Introduction
11.2 Production methods for the important insect pathogenic fungi
11.2.1 Lagenidium giganteum
11.2.2 Leptolegnia chapmani
11.2.3 Coelomomyces spp. Keilin
11.2.4 Entomophthorales
11.2.5 Microsporidia
11.2.6 Ascomycetes Hypocreales
11.2.6.1 Solid substrate fermentation
11.2.6.1.1 The end products of solid substrate fermentation
11.2.6.1.2 Substrates and media
11.2.6.1.3 Equipment for fermentations
11.2.6.1.4 Fermentation parameters: (inoculum, moisture, temperature, aeration, pH)
11.2.6.1.5 Downstream processing
11.2.6.1.6 Major technical problems/solutions in solid substrate fermentation
11.2.6.2 Submerged fermentation
11.2.6.2.1 The end products of submerged fermentation
11.2.6.2.2 Media
11.2.6.2.3 Fermentation parameters (air, agitation, pH)
11.2.6.2.4 Equipment for submerged fermentation
11.2.6.2.5 Downstream processing
11.2.6.2.6 Major technical problems/solutions in submerged fermentation
11.2.6.3 Other, novel, production methods
11.2.6.4 Other ascomycetes
11.2.6.4.1 Cordyceps (Isaria)
11.2.6.4.2 Akanthomyces (Lecanicillium)
11.2.6.4.3 Hirsutella thompsonii
11.2.6.4.4 Metarhizium (Nomuraea) rileyi
11.2.6.4.5 Aschersonia
11.2.6.4.6 Culicinomyces
11.3 Process and quality control in mass production
11.4 Current knowledge about the effect of cultural conditions on propagule attributes
11.4.1 Age of conidia
11.4.2 Conidia produced under certain nutrient conditions or under osmotic stress
11.4.3 Conidia produced after photoirradiation during vegetative growth
11.5 The challenge in mass production of entomopathogenic fungi
References
12 Commercial production of entomopathogenic bacteria
12.1 Introduction
12.2 Biology of commercial entomopathogens
12.3 Pathogenesis and pest control impact
12.4 Culture selection and maintenance
12.5 Inoculum preparation
12.6 Fermentation
12.7 Recovery and concentration steps
12.8 Formulation
12.9 Formulation standardization
12.10 Quality assurance methods
12.11 Conclusion
References
13 Production of entomopathogenic viruses
13.1 Introduction
13.1.1 General introduction
13.1.2 Entomopathogenic viruses
13.1.3 Baculoviruses
13.1.3.1 Taxonomy
13.1.3.2 Baculovirus phenotypes and their function
13.1.3.3 Occlusion-derived virus–midgut interactions
13.2 In vivo production of baculovirus-based biopesticides
13.2.1 Introduction
13.2.2 Increased adoption of nucleopolyhedrovirus products
13.2.3 Production using infected larvae
13.2.4 Challenges for existing baculovirus pesticides and the case for in vitro production
13.3 In vitro production—current status
13.3.1 Introduction
13.3.2 Cell lines available
13.3.3 Virus isolates available
13.3.4 Low-cost media
13.3.5 Current status of bioreactor-based production—HearNPV as a case study
13.3.5.1 Overview of bioreactor-based processes
13.3.5.2 Maximum fed-batch yields reported
13.3.5.3 Economic feasibility
13.4 Limitations to bioreactor production of baculovirus-based pesticides
13.4.1 Lack of a chemically defined media
13.4.2 Low budded virus titers
13.4.3 Occlusion-derived viruses produced in cell culture may have a lower speed of kill
13.4.4 Viral genome instability during in vitro passaging
13.4.4.1 Natural virus populations
13.4.4.2 Selection and generation of mutants in cell culture
13.4.4.3 Instability through transposable elements
13.4.5 Complications with high-density cell culture
13.5 Future research directions for bioreactor production of baculovirus-based pesticides
13.5.1 Chemically defined media for insect cell culture
13.5.2 Genomics/transcriptomics of insect cell lines
13.5.3 Metabolomics of insect cell lines
13.5.4 Genetically modified viruses
13.5.4.1 Viruses with increased speed of kill
13.5.4.2 Viruses with increased infectivity
13.5.4.3 Changing the ratio of budded virus and occlussion-derived virus production
13.5.4.4 Stabilized genomes
13.5.5 Future potential
13.6 Conclusion
Acknowledgements
References
14 Formulations of entomopathogens as bioinsecticides
14.1 Introduction
14.1.1 Goals and benefits of formulations
14.1.2 Challenges of microbial pesticides
14.2 Biological considerations
14.2.1 Biological attributes for the microbe
14.2.1.1 Activity
14.2.1.2 Viability and storage stability
14.2.1.3 Residual activity
14.2.1.4 Efficacy
14.2.1.5 Integration with production
14.2.2 Potential hazards
14.2.2.1 Contamination issues
14.2.2.2 Biohazard worker exposure concerns
14.2.2.3 Allergenicity
14.2.2.4 Combustion, thermal degradation, dust explosion hazards
14.3 Physical considerations
14.3.1 Cost
14.3.2 Formulation form
14.3.3 Ingredients
14.3.4 Processing
14.3.5 Mixing/handling/packaging
14.3.6 Consumer esthetics
14.3.7 Application
14.4 Additional considerations on formulation
14.4.1 Sources of technologies
14.4.2 Legal requirements
14.4.3 Current effective formulations
14.4.4 Unique applications
14.5 Conclusions and future of biopesticide formulations
USDA disclaimer
References
15 Mass production of entomopathogens in less industrialized countries
15.1 Introduction
15.2 Issues and opportunities for entomopathogen uptake in less industrialized countries
15.3 Practical constraints for entomopathogen uptake in developing countries
15.4 Production of entomopathogens in less industrialized countries
15.5 Production of entomopathogenic fungi
15.5.1 The LUBILOSA system
15.5.2 The Caroni system
15.6 Additional examples from other countries
15.6.1 China
15.6.2 India
15.6.3 Brazil
15.6.4 Cuba
15.6.5 Honduras
15.6.6 Kenya and South Africa
15.7 Other systems
15.8 Mass production of baculoviruses
15.8.1 Country case studies
15.8.1.1 China
15.8.1.2 India
15.8.1.3 Thailand
15.8.1.4 South Africa
15.8.1.5 East Africa (Kenya and Tanzania)
15.8.1.6 South America and potato tuber moth
15.9 Other production systems
15.10 Generic production issues
15.10.1 Product quality
15.10.2 Scale of production and application rates
15.10.3 Safety
15.10.4 Economics of production
15.11 Requirements for establishing biopesticide industries in less-industrialized countries
15.11.1 Research and information
15.11.2 Registration and regulation in less-industrialized countries
15.11.3 Responsibility
15.11.4 Future
Acknowledgments
References
Section III
16 Potential and challenges for the use of insects as feed for aquaculture
16.1 Introduction
16.2 Insects in aquafeeds: performances and digestibility
16.2.1 Insect proteins: effects on fish meal and soybean meal sparing
16.2.2 Insect fat and oils: effects on fish and soybean oil sparing
16.3 Insects and fish health
16.3.1 Gut morphology
16.3.2 Immune response
16.3.3 Oxidative status
16.3.4 Gut microbiota
16.4 Challenges and future perspectives
16.5 Conclusions
References
17 The role of insects for poultry feed: present and future perspective
17.1 Introduction
17.2 General nutrient composition of insects and insect-derived ingredients
17.2.1 Impacts of processing method and form
17.2.2 Functional aspects of insects in poultry diets
17.2.2.1 Antimicrobial peptides
17.2.2.2 Chitin
17.2.2.3 Lauric acid
17.3 Insects in meat bird production
17.3.1 Broilers
17.3.2 Other meat birds
17.4 Insects in egg layer production
17.5 Impact of insect-derived ingredients on behavior and welfare
17.6 Barriers and hurdles for use of insects in poultry diets
17.6.1 Organic classification
17.7 Summary and the conclusions
References
18 Insects as food for insectivores
18.1 Introduction
18.2 Nutrient content of insects
18.2.1 Protein and amino acids
18.2.2 Fats and fatty acids
18.2.3 Carbohydrates
18.2.4 Fiber and chitin
18.2.5 Minerals
18.2.6 Vitamins and carotenoids
18.2.6.1 Vitamin A
18.2.6.2 Vitamin D
18.2.6.3 Vitamin E
18.2.6.4 B-vitamins
18.2.6.5 Vitamin C
18.2.7 Other nutrients
18.2.7.1 Choline
18.2.7.2 Taurine
18.2.7.3 Sterols
18.3 Effects of insect size/life stage on nutrient composition
18.4 Effects of insect diet on insect nutrient composition
18.5 Effects of environment on insect composition
18.5.1 Temperature
18.5.2 Humidity
18.5.3 Photoperiod
18.6 Nutrient requirements of insectivores including nutrient availability
18.6.1 Availability and digestibility
18.7 Enhancing the nutrient composition of insects as food for insectivores
18.7.1 Gut loading
18.7.2 Dusting
18.7.3 Feeding nutrient enhanced diets during growth
18.7.3.1 Fatty acids
18.7.3.2 Calcium
18.7.3.3 Carotenoids
18.7.3.4 Vitamin E
18.8 Other considerations
18.8.1 Pathogens/parasites
18.8.2 Heavy metals
18.8.3 Mycotoxins
18.8.4 Other toxins
18.8.5 Uric acid
18.9 Conclusions
References
19 Production of solitary bees for pollination in the United States
19.1 Introduction
19.2 The alfalfa leafcutting bee
19.3 The alkali bee
19.4 The blue orchard bee
19.5 Other solitary bees of interest for pollination
19.6 Concluding remarks
Acknowledgments
References
20 Production of bumblebees (Hymenoptera: Apidae) for pollination and research
20.1 An introduction to rearing bumblebees
20.2 Bumblebee lifecycle
20.3 Pathogens, parasites, and pests—an overview
20.4 Rearing facilities
20.4.1 General setup and equipment
20.4.2 Environmental conditions
20.4.3 Bumblebee rearing units
20.5 Nutrition
20.5.1 Nectar substitute
20.5.2 Pollen provisions
20.5.3 Pollen preparation
20.6 Gyne collection and transportation
20.7 Installing gynes and stimulating broodiness
20.8 Colony care and senescence
20.8.1 Sanitation
20.8.2 Deploying colonies into the wild
20.9 Mating trials
20.10 Overwintering gynes
20.11 Closing remarks
References
21 Current and potential benefits of mass earthworm culture
21.1 Introduction
21.1.1 Ecological groupings
21.1.2 Selection of species
21.1.3 Cultivation techniques
21.2 Current applications
21.2.1 As a protein source
21.2.2 In organic waste management
21.2.3 As fishing bait
21.2.4 In soil restoration
21.2.5 In agro-ecosystems
21.2.6 In laboratory experimentation
21.2.6.1 Moisture
21.2.6.2 Temperature
21.2.6.3 Substrate
21.2.6.4 Feed
21.2.6.5 Density
21.2.6.6 Species interactions
21.2.7 In ecotoxicology
21.3 The future for mass earthworm culture
References
Index
Back Cover